Filtering in pre-evacuated containers

ABSTRACT

An independent blood filter device depends on flow geometry to deliver blood serum or plasma free of detrimental levels of hemoglobin. It depends critically on an upstream flow rate or pressure differential limiting control element or device that limits the rate of change of pressure differential across the filter element. Pre-evacuated versions can be used to simultaneously draw blood from a living being and provide pressure differential across the filter element between an evacuated collector and a supply end open to atmosphere. A unit pressurized by hand motion employs the external shape of a partially filled blood collection tube as a piston to produce pressure in advance of the control element or device to create the pressure differential across the filter element to a collector vented to atmosphere. The control element or device is disclosed in numerous forms, including specially sized flow constrictions and compliant arrangements.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation of U.S. application Ser. No.13/829,424, filed Mar. 14, 2013, now U.S. Pat. No. 9,427,707, whichclaims priority to U.S. Application Ser. No. 61/681,823, filed on Aug.10, 2012. The above-mentioned applications are hereby incorporated byreference.

TECHNICAL FIELD

This invention relates to extraction of fluid of desired characteristicsfrom a small fluid sample, to isolating relatively large particles froma small sample, and to performing assays and similar activities with theseparated substances.

This invention relates specifically to rapid, convenient, inexpensiveand sterile extraction of blood plasma, blood serum and other fluid froma small sample of whole blood. It also relates to isolation of bloodcells and other components from a small sample, and to using smallquantities of a blood-derived, filtered fluid at natural or dilutedconcentrations to perform bio-array assays and other activities such asdiagnostic and analytical procedures. In respect of source of blood tobe used, the invention is highly useful in directly drawing blood, andalso is highly useful with fresh blood previously drawn within a typicalcollection tube or otherwise, and with stored blood that has beentreated fresh to prevent agglutination.

As used here, “Blood Plasma” refers to the liquid component of wholeblood constituting about one half of the volume of the blood, bloodcells constituting the remainder of the volume. “Blood Serum” refers tothe liquid component of whole blood from which blood cells and bloodplatelets have been removed.

BACKGROUND

As traditionally conducted, a set of adult blood tests necessitatesdrawing whole blood with 3 to 6 of the well-known pre-evacuated bloodcollection tubes (e.g. Vacutainer™, Becton Dickinson and Company, EastRutherford, N.J.), each with typically 2 to 10 milliliter capacity.Plasma or serum is typically obtained when whole blond collected in thisfashion is processed by centrifuging or filtering, performed withinminutes from the sample being drawn unless a stabilizing substance hasbeen added to permit delayed separation.

The availability of sensitive biological assays has also made itpossible to run accurate tests employing much smaller sample volumesthan previously employed. For instance, multiple tests are availablethat can be performed employing less than 0.1 milliliter of the fluid,using bio-array techniques. No very simple, inexpensive and rapidlyoperable device has been commercially available for providing serum orplasma extraction at this size volume.

Typical delays in obtaining plasma or serum can range from 10 minuteswhen a centrifuge is on site to over one hour when it is within thefacilities. The delay can be days if samples must be transported toremote locations. These delays defeat the value of onsite diagnosticsmade possible by the new bio-array (biochip) technologies. The majorbenefit of biochip technology is to offer a diagnosis within 15 to 60minutes, saving critical time for intervention as well as saving costs.

Small volume whole blood collection, per se, however, has long beenavailable. It was originally developed for blood tests for infants andsmall animals. For this purpose, evacuated collection tubes have beenavailable for drawing a fraction of a milliliter or a few milliliters ofblood. (Extremely small blood volumes have also traditionally beenobtained by use of a puncture wound. The finger for instance is prickedwith a lancet and then squeezed until a fluid drop of, for example,10-20 microliters is obtained).

In general, current methods for achieving small volumes of serum fromwhole blood typically involve numerous steps and employ multiple piecesof equipment and disposable items. Kits are available for these purposesfrom many sources, examples being: Unopette® (Becton Dickinson andCompany); Fisherbrand® microhematocrit and capillary tubes (FisherScientific Company, Hampton N.H.); and StatSampler® capillary bloodcollection kit (StatSpin, Norwood, Mass.). Each of these relies onmultiple separate components for performing the functions of samplecollection, processing, and recovery.

There have been many attempts to develop more convenient devices, but noreliable, simple and simply-operated hand-held filtering device isavailable that can produce hemolysis-free serum or plasma.

Prior art in the general field include U.S. Pat. Nos. 2,460,641;3,814,258; 4,343,705; 4,477,575; 4,540,492; 4,828,716; 4,883,068;4,906,375; 4,960,130; 5,030,341; 5,181,940; 5,308,508; 5,364,533;5,413,246; 5,471,994; 5,555,920; 5,681,529; 5,683,355; 5,759,866;5,876,605; 5,919,356; 5,979,669; 5,996,811; 6,045,699; 6,170,671;6,261,721; 6,225,130; 6,406,671; 6,410,334; 6,465,256; 6,471,069;6,479,298; 6,497,325; 6,506,167; 6,516,953; 6,537,503; 6,659,288;6,659,975; 6,755,802; 6,803,022; 6,821,789; 7,070,721; 7,153,477;7,767,466; 7,744,820; 7,927,810; and 7,993,847; and US 2010/0093551.

It is recognized to be desirable to work quickly and efficiently withblood samples of the order of 1 milliliter volume. Most proteinanalyzers for instance require 10 to 100 micro-liters per test and it iscommon to employ 10 or so tests. Multiplexed biomarker cassettes, e.g.those employing micro arrays, typically run 8 to 12 assayssimultaneously, and call for less than 100 micro-liter of serum orplasma for the set of assays.

Devices and techniques made possible by the present disclosure cansimply, inexpensively and rapidly meet the need for obtaining suitableblood serum and other blood-derived fluids from small volume whole bloodsamples. Neither centrifuge separation nor other inconvenient techniquesare employed, while sterile separation at point of collection or pointof patient treatment can be achieved.

The level of hemolysis, the presence of hemoglobin within the plasma orserum as a result of cell damage, may not interfere with most diagnostictests and specifically most protein or ELISA tests, but excess hemolysiscould be indicative of patient health conditions that would need to beconsidered, and consequently lead to an erroneous diagnosis. Morespecifically the presence of hemoglobin in serum may yield erroneousreading of the blood potassium concentration. For these reasonsdesirable hemolysis quantifications of low value have been established.

Consequently, in order to be practical, a plasma or serum extractionprocessor device needs to keep damage to red cells to a minimum.

It is important to consider further that the venous puncture commonlycauses some red cells breakage so that the standards that have beenestablished to define levels of acceptable hemolysis leave little roomfor additional hemolysis by serum separation features. This is whereprevious devices have failed to meet exacting standards.

U.S. Pat. No. 4,477,575 teaches the use of glass fibers with diameterfrom 1 to 4 micron can be efficiently used to separate cells from plasmaor serum in a depressurization syringe-like device. The use of this typeof glass fiber has been adopted in later processes as well as thesuction/depressurization serum extraction method, as exemplified by U.S.Pat. No. 5,364,533 that employs pre-evacuation of a device.

Later prior art as exemplified in U.S. Pat. Nos. 7,744,820, 7,927,810and 7,993,847 and US 2007/0082370 describe blood collection and serumseparation using a an internal negative pressure plurality ofinterconnected tubes as well as the use of glass fibers as filtermedium. This prior art attempts to control hemolysis by stratificationof filtration porosity using a membrane with a void ratio under 30%and/or altered retention properties of the filtration column media.

U.S. Pat. No. 5,876,605 similarly uses glass fiber and seeks to minimizehemolysis with suitable mixing of the blood with an aqueous solution.

U.S. Pat. Nos. 5,979,669, 5,996,811, 6,045,699 and 6,170,671 also useglass fiber as a filtrate material and incorporate means to regulateoutflow of filtrate in order to accommodate variation in hematocrit andcontrol hemolysis. They all show how a number of interconnected tubulardevices create a pressure difference by connection to a suction pump ordevice. Typically the final outlet filter membrane is constructed toregulate serum outlet flow.

U.S. Pat. No. 5,979,669 teaches “In another aspect of the blood filterunit of the invention, a flow area-regulating member is provided on theblood filtering material on the filtrate outlet side which is, ingeneral, the microporous membrane. The flow area-regulating member ismade of liquid-impermeable material, and has an opening having an areasmaller than the blood filtering material thereby regulates so thatfiltrate flows out through the opening. A suitable area of the openingis about 20 to 90%, preferably about 50 to 90% of the blood filteringmaterial area on the filtrate outlet side.”

“The flow area-regulating member can be made by various commercialadhesive tapes, plastic film, thin plastic sheet or the like, andadhesive may be applied to the adhering face of the blood filteringmaterial.”

U.S. Pat. Nos. 5,364,533 and 5,979,669 teach the use of a number ofinterconnected and detachable successions of tubes to create a pressuredifference across a filter assembly in order to obtain plasma byfiltration.

U.S. Pat. Nos. 6,506,167, 6,659,288 and 6,045,699 suggest the use ofstratified filtration column as well as external active sequencing ofcontrolled differential pressure forcing the blood through the filtercolumn or the entire device from blood inlet to filtrate outlet.

U.S. Pat. No. 6,045,699 teaches that a suitably hemolysis-free filterdevice can be constructed where pressure differential across a filterassembly of an evacuated device is actively controlled from a tetheredpressure source external to the filter device. It teaches to sequencethe pressure differential with a pressure sequencer where filtrationbegins with a low pressure differential which is “controllablyincreased” as filtration progresses. The patent teaches using activeexternal equipment such as a peristaltic pump or a syringe. It teachesto “trace” pressure different variation with time and to “adjust suctionor pressurizing speed.”

U.S. Pat. No. 7,993,847 teaches the use of filter assembly in which amembrane exit filter, in a passive warmer, regulates the pressuredifferential across a filter assembly, seeking to yield a substantiallyhemolysis-free serum sample.

The membrane exit filter has a number of micron size apertures. But sucha membrane is totally ineffective to limit the flow of air across it asair molecules are sub angstrom in dimensions. Such a membrane iseffective only to limit liquid flow and have any effect much later inthe filtration process when blood has already reached and serum orplasma has already traveled through the filter assembly. Such a devicestarts the filtration process with maximum pressure differential acrossthe filter assembly and is insufficient to control hemolysis to the lowlevel necessary.

A prior attempt by one of us to meet the present need is shown inUS2010/0093551. It has the requirement of repeated hand movements andother drawbacks, and lacks the critical flow rate or pressuredifferential-limiting element or device geometry now to be described.Like many other attempts to meet the need, has not been commercialized.

SUMMARY

The present invention teaches how to make a totally independent filterdevice with few parts able to induce controlled pressure differentialconditions that permits delivery of suitably hemolysis free serum orplasma from blood. The blood may be undiluted whole blood that issimultaneously drawn from a subject. The blood may be sourced fromanother vessel.

The subject of this invention is to offer a blood filtration method toobtain serum that accommodates hematocrit variations and delivers anacceptable level of hemolysis. This invention contrasts in two ways withprior art. First this invention teaches how to minimize hemolysis bypassive control of the pressure differential forced upon the bloodthrough the filter assembly. Second this invention teaches how tocontrol in a passive way the pressure differential forced upon the bloodthrough the filter assembly by controlling the inflow rate of bloodprior to contact with the filter assembly.

In addition this invention in contrast with prior art, teaches how tobuild such a filter device using a single tube, therefore minimizingmanufacturing costs.

Another aspect of this invention is to offer a method of extracting byfiltration a volume of serum from blood with a minimum of hemolysis.

This invention teaches how to sequence in a totally passive manner thepressure differential across the filter assembly of an evacuated deviceand yield substantially hemolysis-free serum. As blood is introducedinto the device the filtration is caused to proceed with only a slowlyrising pressure differential followed with a very slowly decliningpressure differential and termination of the process. This inventionteaches in a passive manner the control of the pressure differentialacross a filter assembly in an evacuated device through the control ofthe blood intake flow rate. The control of the pressure differentialtakes effect as the blood enters the device in contrast to the teachingof U.S. Pat. No. 7,993,847 where control begins much later and onlyafter a quantity of blood has reached and plasma or serum has traveledthrough the entire filter assembly.

Another aspect of this invention is a mechanically simple method topassively control the magnitude of the pressure differential across bothends of an evacuated hand-held tube-like device separated by a filterelement as blood enters one end as shown on FIGS. 1A and 1B.

Another aspect of this invention is a mechanically simple method topassively control the rate of change of the pressure differential acrossboth ends of an evacuated hand-held tube-like device separated by afilter element as blood enters one end as shown on FIGS. 1A and 1B.

Another aspect of this invention is a mechanically simple method topassively control the magnitude of the pressure differential across bothends of an evacuated hand-held tube-like device separated by a filterelement by controlling the rate of entry of the blood into the device asshown on FIG. 1A.

Another aspect of this invention is a mechanically simple method topassively control the rate of change of the pressure differential acrossboth ends of an evacuated hand-held tube-like device separated by afilter element by controlling the rate of entry of the blood into thedevice as shown on FIG. 1A.

Another aspect of this invention is an evacuated hand-held tube-likedevice holding in its central region a filter element and a flow ratecontrolling element constructed such that as blood enters the device viathe flow rate controlling element the magnitude of the pressuredifferential across the filter element is controlled by the flow ratecontrol element as shown on FIG. 1B.

Another aspect of this invention is an evacuated hand-held tube-likedevice holding in its central region a filter element and a flow ratecontrolling element constructed such that as blood enters the device viathe flow rate controlling element the rate of change of the pressuredifferential across the filter element is controlled by the flow ratecontrol element as shown on FIG. 1B.

The device is intended to be used instead of a common pre-evacuatedblood collection device such as a BD Vacutainer™ and can deliver serumor plasma by filtration directly without use of a centrifuge. Itincorporates a flow rate control section preceding the filter, which maybe internal to the evacuated tube or external. Blood is drawn into thepartially evacuated device and with appropriate flow rate traverses toand through a filter assembly that captures cells but permits serum orplasma to flow through into a collection chamber.

The device enables simple and rapid extraction of blood serum or plasmain milliliter quantities from a collected blood sample. The device canalso provide for the addition of an agent that may coat the filter orthe tube. Syringe extraction of blood serum from the device can beachieved via an access septum located at the downstream end of thecollection tube. The device permits all functions to be performedrapidly, without exposure of personnel to needles, and with minimumdanger of exposure of the operator to the sample or contamination of thesample while enabling standard evacuated collection tube methods to beused.

In preferred implementations, the invention is a blood separation devicein the form of a cylindrical tubular assembly similar in shape to a 6 mlVacutainer™ It incorporates an input flow rate control element and froma drawn blood sample somewhat smaller than 2 milliliter producesapproximately a 0.25 milliliter volume of blood serum practically freeof hemoglobin.

In some preferred implementations the input flow rate control elementmay be internal to the tube-like device.

In other preferred implementations the input flow rate control elementmay be external to the tube-like device.

In some preferred implementations the invention incorporates, within theblood input chamber, an elastically compressible element such as aclosed cell member of resilient plastic or rubber foam or an air-filledbladder that regulates the rate of evolution of the pressure differenceacross the filter assembly as blood enters the region of thecompressible element.

Preferably, neither air nor gas is permitted to enter any part of thedevice until the filtration process has been completed and the serumchamber has been brought to atmospheric pressure by letting air atatmospheric pressure enter through the serum access septum or through anequivalent port. That process takes approximately 1 or 2 minutes.

Another aspect of this invention is a mechanically simple method topassively control the magnitude of the pressure differential across bothends of pressurized hand-held tube-like device separated by a filterelement as blood enters one end as shown on FIGS. 1D and 1E.

Another aspect of this invention is a mechanically simple method topassively control the rate of change of the pressure differential acrossboth ends of an evacuated hand-held tube-like device separated by afilter element as blood enters one end as shown on FIGS. 1D and 1E.

Another aspect of this invention is a mechanically simple method topassively control the magnitude of the pressure differential across bothends of a pressurized hand-held tube-like device separated by a filterelement by controlling the rate of entry of the blood into the device asshown on FIG. 1C.

Another aspect of this invention is a mechanically simple method topassively control the rate of change of the pressure differential acrossboth ends of a pressurized hand-held tube-like device separated by afilter element by controlling the rate of entry of the blood into thedevice as shown on FIG. 1C.

The independent blood filter device depends on flow geometry to deliverblood serum or plasma free of detrimental levels of hemoglobin. Itdepends critically on an upstream flow rate or pressure differentiallimiting control element or device that limits the rate of change ofpressure differential across the filter element. Pre-evacuated versionscan be used to simultaneously draw blood from a living being and providepressure differential across the filter element between an evacuatedcollector and a supply end open to atmosphere. A unit can be pressurizedby hand motion employing the external shape of a partially filled bloodcollection tube as a piston to produce pressure in advance of thecontrol element or device to create the pressure differential across thefilter element to a collector vented to atmosphere. The control elementor device is disclosed in numerous forms, including specially sized flowconstrictions and compliant arrangements.

The features described in the preceding pages are comprehended in thefollowing summary:

In a first aspect, the invention features a filtering device forfiltering blood to obtain serum or plasma in a container, the containerhaving access at both ends, a filter located within the container, and aflow rate or pressure differential limiting control element or device,the limiting element or device located upstream of the filter.

Preferred implementations of this aspect of the invention mayincorporate one or more of the following:

The container may be partially evacuated. The limiting control elementor device may be located outside the container. The limiting controlelement or device may be integral with a blood drawing needle assembly.The filtering device may be fitted with an entering flow rate limitingcontrol element or device. The container during operation may bepartially pressurized. The limiting control element or device may belocated inside the container. The limiting control element or device maybe a flow constriction element or device. The flow constriction elementor device may be in the form of a pin hole or pin holes in aflow-blocking disk. The flow constriction element or device may be inthe form of or may comprise a selected length of capillary tubing. Theflow constriction element or device may be in the form of or maycomprise a fine mesh or porous foam. The flow constriction element ordevice may be in the form of or may comprise a passage defined by ascrew like segment. The filtering device may have a limiting controlelement or device constructed to limit differential pressure acrossblood in the inlet side of the filter. The filtering device may have alimiting control element or device preceding the filter that limitsentering flow-rate of whole blood. The filtering device may have alimiting control element or device preceding the filter that definesentering pressure increase rate in whole blood. The filtering device maybe portable or hand held, and may include a volume sized for blood drawnfrom a living being. The material of the filter may comprise glassmicrofibers and micro-porous membrane on a locating support. Thecontainer may be a tube. The filtering device may have an access septumat the inlet end of the container or tube. The filtering device may havean access septum at the outlet end of the container or tube. Thefiltering device may have, at the outlet end of the container or tube, aremovable element in the form of an end-plug with a serum or plasmaholding cavity. The filtering device may include a volume pre-evacuatedfor drawing blood from a source. The volume may be pre-evacuated fordrawing blood from a living being. The filtering device may beconstructed for controlling the incoming blood flow rate into acontainer or tube holding a filter in its central region such that therate of increase of the pressure differential between the two sides ofthe filter stays below 30 mmHg per second. The filtering device may beconstructed to limit rate of increase to stay below 20 mmHG per second.The filtering device may be constructed, by the axial position of thefilter in the container or tube, for controlling the rate of increase ofthe pressure differential between the sides of the filter to stay below30 mmHg per second. The filtering device may be constructed to limitrate of increase of pressure differential to stay below 20 mmHg persecond. The filtering device may be pre-evacuated to induce flow ofwhole blood into the device, and may be constructed to define incomingblood flow rate into the volume at the inlet side of the filter toincrease the pressure differential across the filter at a rate below 30mmHg per second. The filtering device may be constructed to limit rateof increase of pressure differential to stay below 20 mmHg per second.The container may be a tube and the pressure differential may be betweenthe ends of the tube. The limiting control element or device may be aninsertion of a compressible closed cell volume. The filtering device,following blood collection, may be constructed to be-pressurized bymanual action of the user to produce pressure on collected blood toforce the blood through the control element or device, and through thefilter, to a vented collector. The filtering device may be adapted foruse with a first tubular member which is a pre-evacuated bloodcollection member, and the device may comprise a member pre-fitted inshape to receive the first tubular member and to be moved relative tothe first tubular member to produce positive pressure, preceding aninternal whole blood flow constriction element.

The pressure differential across the filter may be limited to below 30mmHg per second. The pressure differential across the filter may belimited to below 20 mmHg per second. The flow rate through the filtermay be approximately 2 to 10 cc per minute. The flow rate may be between3 to 6 cc per minute. The device may be constructed to produce a volumeof between about 1 to 2 cc filtrate. The device may be constructed toproduce a volume of about 1.5 cc filtrate. The limiting control elementor device may be a tubular element between ½ inch and 4 Inches in lengthand may have an internal diameter between about 0.008 and 0.013 inch.

In another aspect, the invention comprises a method of obtaining bloodserum or plasma using the filtering device according to the first aspectdescribed alone or together with any of the further features mentioned.

The major benefits offered by the devices are:

-   -   Cost saving    -   Simplicity of operation    -   Under three minute plasma delivery    -   Serum availability at the point of care    -   Protection of the operator from exposure    -   Freedom of contamination of the sample    -   Elimination of the need for a centrifuge

The details of one or more embodiments of the invention are set forth inthe accompanying drawings and the description below. Other features,objects, and advantages of the invention will be apparent from thedescription and drawings, and from the claims.

In FIGS. 1-6 the geometry of a flow rate or pressure differentiallimiting control element or device upstream of the filter is used todefine conditions with pre-evacuated tubes. The tubes may be bloodcollection tubes. In FIGS. 7-9 the geometry is used in respect of apressurized system.

DESCRIPTION OF DRAWINGS

FIG. 1 illustrates prior art;

FIG. 1A diagrammatically indicates the flows of devices according topresent invention having a flow regulator upstream of a pre-evacuatedcollection device containing a filter assembly, the device shown havinga tubular housing;

FIG. 1R similar to FIG. 1A, indicates the flows of devices having a flowor pressure regulator within a pre-evacuated collection device, upstreamof a filter assembly within the device, the device shown having atubular housing;

FIG. 1C diagrammatically indicates the flows of devices according topresent invention having a flow regulator upstream of a pressurizedcollection device containing a filter assembly, the device shown havinga tubular housing;

FIG. 1D similar to FIG. 1C, indicates the flows of devices having a flowor pressure regulator within a pressurized collection device, upstreamof a filter assembly within the device, the device shown having atubular housing

FIG. 2 is an implementation of the device generically illustrated inFIG. 1B, the device having a capillary flow regulator between first (A)and second (B) blood holding chambers upstream of the filter assemblywithin a tubular housing and a needle-penetrable access septum at theend of the tube for serum or plasma;

FIG. 2A is an implementation of the device generically illustrated inFIG. 1B, the device having a capillary flow regulator between first (A)and second (B) blood holding chambers upstream of the filter assemblywithin a tubular housing and a sealed but removable slide-fit end plugthat defines a holding chamber for serum or plasma;

FIG. 2B is an implementation of the device generically illustrated inFIG. 1B, the device having a capillary flow regulator between first (A)and second (B) blood holding chambers upstream of the filter assemblyand a sealed but removable end holding chamber for serum or plasma, thatis sealed and held to the tubular housing by a bayonet coupling device;

FIG. 2C is an implementation of the device generically illustrated inFIG. 1A, the device having a flow regulator upstream of the entry to ablood holding chamber preceding the filter assembly and a sealed butremovable end holding chamber for blood serum or plasma, that is sealedand held to the tubular housing by a bayonet coupling device, the figurefurther illustrating the arm of a human subject (reduced scale) andusual blood collection needle and connection tubing for conducting bloodfrom the subject;

FIG. 2D is an implementation of the device generically illustrated inFIG. 1B, the device having a flow regulator in the form of a narrowscrew-thread-defined helical passage, between first (A) and second (B)inlet holding chambers upstream of the filter assembly and a sealed butremovable end holding chamber for serum or plasma, that is sealed andheld to the tubular housing by a bayonet coupling device;

FIG. 2E is an implementation of the device generically illustrated inFIG. 1B, the device having a pin hole flow regulator passage between ablood holding chamber upstream of the filter assembly and a sealed, butremovable, end holding chamber for serum or plasma, that is sealed andheld to the tubular housing by a bayonet coupling device;

FIG. 2F is an implementation of the device generically illustrated inFIG. 1B, the device having a resiliently collapsible flow regulator(e.g. air-filled bladder or mass of collapsible rubber-like foam)bladder within an inlet blood holding chamber upstream of the filterassembly and a sealed, but removable, end holding chamber for serum orplasma, that is sealed and held to the tubular housing by a bayonetcoupling device;

FIG. 3 is a photograph of a blood collection and flow regulator assemblyand separate insertion guide for use with a device according to FIG. 1A,while FIG. 3a illustrates the details of an example of the flexibletubing and external flow regulator to be disposed upstream of thecollection/filter unit portion of the device;

FIG. 4 is a side cross-section view and FIG. 4′ a top view of animplementation of a cup-shaped holder for the capillary flow regulatorelement useful in implementations according to FIGS. 2, 2A, and 2B;

FIG. 4A is a side cross-section of a helical path flow regulator(constrictor) formed by a screw thread inside the tubular housing of thedevice of FIG. 2D while FIG. 4A′ is a side view of thescrew-thread-defining element;

FIGS. 5 and 6 are pressure vs. time plots of the pressures within apre-evacuated blood collection device with internal filter assemblyrespectively with no flow control, and the flow control of the deviceaccording to FIG. 1A employing the flow regulator of FIG. 3A;

FIG. 7 is a longitudinal cross-section of a filter device assembly and acollection tube holding blood in position to be inserted into the filterdevice, here the collection tube shown is fitted with an access septum;

FIG. 7A shows a collection tube in process of being inserted into thefilter device;

FIG. 8 is a longitudinal cross-section of a filter device assembly and acollection tube holding blood fully inserted into the filter device andfollowing opening the “Serum Holding Chamber” to atmospheric pressure;

FIGS. 9A, 9B and 9C show blood-transfer-flow regulator devices locatedabove the glass fiber filter;

Like reference symbols in the various drawings indicate like elements.

DETAILED DESCRIPTION

In the presently preferred implementation the device comprises atube-shaped assembly closed at each end with a needle-penetrable accessseptum. The blood inlet access septum located at one end of the tubeconnects for inlet of blood to the blood holding chamber and the outletaccess septum located at the other end of the tube faces theserum/plasma collection chamber and can function as the air inlet portto terminate the filtration process. A filter assembly is fixed in placein the central region of the tube. A passive blood flow controllingsegment may be located between the blood inlet access septum and thefilter assembly (FIG. 1B) or may be external, preceding the device (FIG.1A). FIGS. 1A and 1B illustrate flow geometries based on pre-evacuationof container or tubes. Preferred implementations are shown in FIGS. 2-6.FIGS. 1C and 1D illustrate similar flow geometries based onpressurization of the container or tube, similar preferredimplementations of which are shown in FIGS. 1-9.

The inlet access septum is adapted for being pierced by a standardblood-collection needle assembly (needle penetrable) and defines one endof the chamber free to accept the blood sample for filtration. A flowrate regulating segment adjacent to this access septum regulates therate of flow of blood approaching the filter assembly and defines thepressure differential driving the filtration process in thepre-evacuated unit. The filter assembly is preferably designed to coverthe entire cross-section area of the tube. The filter assembly capturesthe cellular components of the blood and permits passage of the serum orplasma components. The filter assembly preferably terminates with aperipherally sealed element that prevents flow-around (bypass flow) ofblood product and an axial retainer pressed or molded into place.

The axial location of the filter assembly and starting point pressure(vacuum) level of the pre-evacuated device can be used to coordinate thepressure differential changes across the filter assembly.

A volume of elastomeric compressible media or a resiliently collapsibleelement may be located in the chamber free to accept the blood sample asshown on FIG. 2F for modulating the pressure differential. Preferablyclosed cell silicone sponge or foam made of natural rubber, or Nitrile,with durometer less than Shore 45 may be used, or a partially air-filledbladder, for instance.

The terminal end of the tube forms the low pressure chamber that inducesthe filtration process and is the serum collection chamber. It is closedwith a second access septum through which atmospheric air can be causedto enter to equilibrate pressure across the filter assembly to terminatefiltration, and for subsequent removal of filtered material via a needleand syringe.

In its presently preferred implementations shown on FIG. 2 the serumcollection access septum has a hollow space sized to hold all thefiltrate and can be slide-ably removed from the tube for serumaspiration with a pipette.

In another implementation the scrum collection closure segment can berigid and held in place with a simple pressed in O ring as shown on FIG.2A or be detachable via a bayonet connector as shown on FIG. 2B toimplement depressurization and access to the filtrate.

In preferred implementations a region adjacent to the blood inlet accessseptum of the tube is dedicated to hold the blood sample to be filtereduntil filtration has been performed and to retain all extraneous bloodand blood components, liquid and gaseous.

One aspect of the invention is the incorporation of an intake blood flowrate controller/regulator prior to the filtration stage, FIG. 1A. Incertain preferred embodiments the controller/regulator of the rate ofblood flows is located within the device, FIG. 1B.

In certain preferred embodiments the flow rate controller/regulatorlimits the rate of blood flow that can enter the filter assembly. FIG. 2and FIG. 4b In another embodiment the controller/regulator/restrictor islocated between the access septum and the filter assembly as shown onFIG. 2D; FIGS. 4A and 4A′ show details of the restrictor (constrictor).

In another implementation the flow rate regulation function can beimplemented externally from the tube-shaped assembly, FIG. 1A. Apreferred implementation is shown on FIG. 2C and incorporated into theblood delivering needle assembly, FIGS. 3 and 3A.

In various implementations the flow rate regulating function can beimplemented at the inlet of the filter assembly or within the filterassembly or a combination of both.

Preferred implementations have one or more of the following features:

The interior of the tube assembly may be evacuated by inserting theaccess septum to close the upstream tube as the assembly is in alow-pressure chamber or by piercing the installed access septum with aneedle connected to a vacuum pump. It is expected that vacuum can bemaintained for a minimum of one year.

The device incorporates a filter or filter material assembly to whichblood entering the upstream tube is exposed. In preferred embodimentsthe filter assembly may have 3 constituents:

-   -   A first component that promptly disperses the blood across the        entire section of the filter assembly. This is preferably a        highly hydrophilic, highly porous material such as Porex filter        material POR 410 or POR 4711. In another construction the upper        layer of the next filter element can be conditioned to perform        this function.    -   A second filter element, a suitable thickness of glass fiber        filter material such as Johns-Manville Micro-Strand Glass        Microfibers with diameter between 1 and 4 micron and packed in        density between 0.2 and 0.5 g/ml. preferably the thickness is        between 10 and 20 mm.    -   A third component is a micro-porous membrane able to block        passage of cell debris as well as glass fiber debris and        preferably permits passage of particles or molecules smaller        than 0.6 micron such as plasma or serum. It also serves to        prevent flow around the filter assembly and is sealed to the        tube on its axial periphery via a compression ring pressing        axially against a ledge internal to the tube. This third        component is preferably a compliant filter material        approximately ½ mm thick such as can be obtained from T.W.        Tremont. Other seal methods may be used such as bonding, thermal        bonding and ultrasonic welding.

The filter assembly is retained and supported axially near the middle atthe tube with a perforated screen member. Suitable glass fiber densityis maintained by axial compression against such screen member. Thesection of the tube between the input access septum and the filterassembly offers a holding chamber for the incoming blood before ittravels through the filtering material.

It is thought that the low-density glass fiber filter material catchesblood cells gradually by entangling at first large blood cell componentsand then smaller blood cell components in the space structure whilepermitting smaller molecules to travel through.

The invention teaches to deliver cells into and through the filterassembly with minimum and controllable force derived from a controlledpressure differential between the blood entering the filter assembly andthe serum collection section of the tube. The pressure differential iscontrolled to induce a low velocity of the blood components beginning atthe initial stage of filtering to minimize shear force on the cells, orimpelling damage from collision with glass fibers of the filter assemblyor with cell lodged in a tangle of glass fiber, in a manner to avoidexcess hemolysis.

We know that red cells are robust when subjected to substantial pressurevariations, but are very fragile in shear. This may explain why a slowerflow rate reduces hemolysis. One other explanation is that red bloodcells can burst on impact with the glass fibers of the filter and thatthe impact damage can be reduced or eliminated if the inrush speed iskept low enough. There is also possibility that cell damage is caused bya high pressure differential across the glass fiber filter, whichsqueezes the red cells in an extreme shear condition into the smallerfilter channels causing greater shear stress that bursts the cells. Inthe latter instance, the longer a high pressure differential exists, themore red cell damage would occur. FIGS. 5 and 6 show that a highpressure differential persists substantially longer when the inflow rateis higher. It has been noticed during experiments that introducing asudden high pressure differential by removing the blood inlet septum andexposing the filter inlet side to atmospheric pressure invariablyresulted in an unacceptable amount of hemolysis, and so, whatever thecause or causes of red cell damage, excess pressure differential must beavoided.

This is achieved by proper dimensioning of the blood receiving volume,the flow rate controlling device (or devices) the volume and density ofthe glass microfibers, the total volume of the tube as well as theinitial level of depressurization of the device, optimization to befound by a series of reasonable trials.

The present invention also teaches to deliver blood in a condition whereearly in the blood injection process a barrier is established betweenthe parts of the tube on either side of the filter assembly. Bloodentering the intake region of the filter assembly diffuses rapidlythrough the hydrophilic media and creates an air tight seal.Consequently the pressure condition in the tube downstream of the filterassembly is little altered by the blood injection. In contrast thepressure within the segment of the tube upstream from the filterassembly is substantially raised by the introduction of blood. Thiscondition creates a pressure differential across the filter assemblythat propels the small molecules contained in the serum to travelthrough the filter assembly.

The invention teaches how to regulate the pressure differential acrossthe filter assembly. This is best achieved by control of the rate ofinflow of blood as it alters the pressure in the tube upstream from thefilter assembly and more specifically the region of the tube in directcontact with the filter assembly. The filter assembly is in cooperativerelationship with the blood which diffuses readily through it by surfacetension as well as pressure differential. Hemolysis takes place as theblood travels through the filter and is strongly affected by thepressure forces and rate of flow through the filter assembly. Little ifany hemolysis takes place as blood enters the intake reservoir, it isthought, based on voluminous experience with Vacutainer™ type devices.

The pressure within the tube is altered by the introduction of thevolume of blood. Considering the Ideal Gas Law:

PV=nRT

where P is the pressure of the gas, V is the volume of the gas, n is theamount of substance of gas (also known as number of moles), T is thetemperature of the gas and R is the ideal, or universal, gas constant,equal to the product of Boltzmann's constant and Avogadro's constant.

In SI units, n is measured in moles, and T in Kelvin. R has the value8.314 J·K⁻¹·mol⁻¹ or 0.08206 L·atm·mol⁻¹·K⁻¹.

Assuming constant temperature, typically human body temperature, theequation simplifies to:

PV=Constant

Initial depressurization of both ends of the tube assembly may be from250 to 700 mmHg. Atmospheric pressure is typically 760 mmHg. Blood, uponwetting the intake side of the filter media, establishes a gas-tightsurface barrier almost immediately, preventing air exchange transportbetween the two ends of the tube. Measurements show that about 0.5 ccare sufficient to form a seal: this occurs within 6-8 seconds when flowrate is kept low enough to prevent hemolysis, and within 1-2 seconds athigher flow rates. Thus, if the blood continues to enter at a high rateof flow the trapped air is compressed and the pressure risesaccordingly. The pressure in the tube upstream from the filter assemblycan rise to near atmospheric pressure while the downstream pressureremains low. This causes a high-pressure differential across the filterassembly, and red blood cells are forcefully pushed into the glassfibers, causing hemolysis. This is a condition analogous to opening theaccess septum to atmospheric pressure after blood injection; it is knownthat this results in a high level of hemolysis. This pressure conditionis exemplified on FIG. 5 showing an average initial pressuredifferential rate of 56 mmHg/sec.

A slow rate of entry of the blood into the tube allows time for theblood to start passing through the filter media; trapped air will stillbe compressed by the incoming blood though much less so, resulting in asmaller pressure differential across the filter and thus minimalhemolysis. This pressure condition is exemplified on FIG. 6 showing anaverage initial pressure differential rate of 13.3 mmHg/sec.

Intake blood flow rates, initial pressure conditions, volumes of bothsegments, upstream and downstream from the filter assembly as well asthe proper filter construction can be optimized to accommodate the rangeof plasma viscosity encountered in practice.

As the filter assembly is terminated with a submicron porosity media,the total volume of blood intake is limited to the free space upstreamfrom the filter assembly less the volume of the filter material takinginto consideration the serum filtered into the downstream tube. Theserum filtration process is self-limiting and brief, 15 to 30 secondstypically.

Using this blood-collecting tube it is possible to carry out bloodcollection and separation in an efficient manner by the followingprocedure:

After sticking the blood-drawing needle into a blood vessel (atatmospheric pressure) or a vein (at near atmospheric pressure) theblood-collection needle punctures through the blood inlet access septumof the device. FIGS. 3 and 3 a show a typical blood sampling kit: theblood drawing needle is the one with the batwing device. At this point,blood is drawn into the accumulation segment of the tube due to thenegative pressure within the entire device. Blood will approximatelyfill that segment.

Shortly after blood enters the accumulation segment it propagates withinthe front part of the filter assembly creating a seal that prevents gasmolecules passage through it. The entry of the blood reduces the spaceoccupied by the molecules of air within the device.

At the start of the process, due to pre-evacuation, the entire device isat a low pressure level, possibly 100 mm Hg. The slow blood entry slowlyfills the volume previously available to the air molecules andconsequently the pressure within that space increases slowly accordingto the Ideal Gas Law.

In the preferred embodiment the device is similar to a 6 cc Vacutainer.It has uniform inside diameter of approximately 10.5 mm and wallthickness of approximately 1 mm. The blood entry chamber, flow regulatorand filter assembly has a length of approximately 33 mm and the overalltube approximately 80 mm.

The filter assembly is formed with approximately 0.35 gram of 108 A or108 B Micro-Strand Glass Microfibers from Johns Manville or equivalentwith nominal diameter 1.8 micron having a net density 0.15 and 0.5 andpreferably approximately 0.027 gram per cubic centimeter. (In otherembodiments 0.5 grams of the microfibers can be used, or within the 0.35gram to 0.5 gram range, 0.415 grams bay be used.)

The glass fiber segment may be covered at its entry with a highlyhydrophilic filter layer such as Porex™ filter material POR 41210 or POR4711 and at its exit with a 0.6 micron porosity filter. (In anotherembodiment filter material of 1.0 micron porosity may be used to takeadvantage of better tear properties that it may have.)

A flow control regulator is located between the blood entry accessseptum and the filter assembly segment. It can be a cup shaped thincylindrical element holding in its center a capillary flexible tubingwith 0.25 mm inside diameter and a length of 40 or 50 mm as shown onFIG. 3A.

The rate of blood flow entering the device through the access septum isquite low, approximately 0.05 cc/sec. to 0.1 cc/sec and when the bloodhas approximately filled the accumulation segment the blood-collectingneedle can be disconnected from the access septum in a manner that doesnot permit air or a gas to penetrate the device. This process takes form15 to 30 seconds.

The pressure differential acting on the blood against the filterassembly rises slowly in a passive manner to approximately 330 mm Hg andsettles to approximately 150 mmHg within 1 to 3 minutes when the serumseparation can be finalized by permitting air at atmospheric pressure toenter the serum end of the tube.

Due to this pressure difference, the blood gains a tendency to flowthrough the flow rate regulation segment and into the filter assemblyand toward the downstream end of the tube. The flow rate regulatorprevents rapid inrush of blood cells and serum molecules. However,because the filter assembly captures cells and only permits throughpassage to molecules or particles smaller than 0.6 micron only serum orplasma or hemoglobin are allowed to pass through and accumulate into thedownstream end of the tube. Thus, separation of the blood is performedshortly after it has been collected.

Upon completion of serum collection, the serum access septum can bepierced or separated for plasma collecting and further processing.

The flow regulator device is preferably in the form equivalent to alength of channel of small cross section (though many times the width ofblood cells). The blood flow rate needs to be such that blood enteringthe glass fiber filter section do not cause damage to the red cellspreviously located in the maze of glass fibers forming the main part ofthe filter. The flow control device permits a steady flow rate andprevents a burst flow from taking place. The process can accommodate theexpected range of blood viscosities.

In another preferred embodiment the flow rate controller is incorporatedin the blood-collection needle assembly and consists of a capillaryrestricted channel approximately 25 to 50 mm long a with diameterbetween 0.25 mm and 0.30 mm.

In another preferred embodiment the blood flow rate controller is in theform of a circular channel created when a screw is inserted in a smoothcylinder of mated diameter. The section of the channel thus created andits length—the number of turns times the diameter-limits the rate offlow possible for a fluid of defined viscosity and a defined pressuredifferential acting on the fluid. Such a screw constrictor is shown onFIG. 2D and FIG. 4A. The channel in the preferred embodiment has asection equivalent to that of a tube of diameter 0.25 mm and 0.30 mm anda length 25 and 50 mm.

Flow Rate and Flow Geometry

The preferred flow rate is from approximately 2 to 10 cc per minute andpreferably 3 to 6 cc per minute to a volume of 1 to 2 cc preferably 1.5cc.

The capillary restriction flow rate for blood can be derived from theHagen-Poiseuille law:

Q=K·ΔP·TrR ⁴/8Lμ

Where:

-   -   K: is a constant    -   Q: flow rate    -   R: capillary radius    -   L: capillary length    -   ΔP: pressure drop    -   μ: blood viscosity

Considering that it is desirable to limit the pressure differential andmaintain a practical flow rate it is possible to select alternate tubediameters and corresponding tube length for either internal or externalflow rate control device or baffle disc, located on the inlet side ofthe filter assembly, fitted with one or numerous pinholes.

If one chooses an equivalent baffle with a single pin hole theHagen-Poiseuille law suggests a 1 mm thick baffle with 0.1 mm diameterhole or a 1/16 inch thick baffle with a 0.005 inch diameter hole asshown on FIG. 2E.

If one might seek to preserve the 12-inch flexible tube length ofcommercial blood drawing assemblies, and accomplish the flow ratecontrol in a blood-drawing implementation, just by special constructionof the tubing, effectively making the tubing itself the limiting controlelement, the Hagen-Poiseuille law instructs that the tubing should havean inner diameter of approximately 0.015 inches, considerably smallerthan that of commercial blood drawing devices. An alternate design isthe introduction of a section of tubing less than the full length of theblood drawing tube that has reduced diameter. According to a preferredimplementation, a 2 to 4 inch section of 0.012 inch diameter tubing isemployed within the 12 inches length from needle to needle, as hereinpresented.

The Hagen-Poiseuille law is applicable to Newtonian fluids. Blood is anon-Newtonian fluid and this is specially expressed when capillaries orrigid flow channels are either too narrow or too long. It has beenverified experimentally that the Hagen-Poiseuille law is useful for thepresent purposes, and is especially applicable to the preferred flowconstrictor, of the order of 0.011 inch internal diameter and 2 incheslength.

It has been verified experimentally that the law does not apply tocapillaries 0.004 or 0.005 inch (100 and 125 micron) in inside diameter.

It has also been verified that extending the length of a rigid tubing to24 inches damages red cell and causes hemolysis.

Preferred dimensions for a tubular limiting control element are betweenabout ½ inch and 4 inches in length and ID between about 0.008 and 0.013inch.

In another way the rate of increase of the pressure differential betweenthe blood entry segment of the device and the serum collection segmentcan be regulated with the insertion of a compressible element working asan intake pressure buffer in the blood entry segment of the device.

In another way the rate of increase of the pressure differential betweenthe blood entry segment of the device and the serum collection segmentcan be facilitated with appropriate volume relationships defined by theaxial location of the filter assembly.

Pressurized Operation

In other uses of control of pressure or flow rate upstream of a bloodfilter using a simple flow rate or pressure control element or sectionas herein described, the pressure differential across the filterassembly is obtainable by pressurizing the blood upstream of the controlelement or section to above atmospheric pressure and venting thedownstream side of the filter assembly to atmosphere.

Highly useful blood separators that implement this approach can make useof a blood container, e.g., a conventional evacuated blood collectiontube, as a novel one-stroke piston to produce the pressure upstream ofthe blood. The blood separator device may take the form of an open endedtube that precedes a filter assembly, into which the blood containerslides. It makes sealed engagement with the tube wall to produce pumpingaction. During this action, the filter assembly and following filtratecollector are closed to the atmosphere. The motion of the container isemployed to increase air pressure throughout the closed volume. Later,upon venting the filtrate collector, the air pressure above the blood inthe container is employed to drive the blood through the control elementor section and filter assembly into the then-vented collector.

Referring to FIGS. 7-9, an implementation is shown in which bloodseparating device 8 is used with a conventional evacuated collectiontube 10 such as available from Becton Dickinson and Company under thetrademark Vacutainer™). When tube 10 is inverted with its rubber accessseal 10 a down, previously collected blood may reach level L, occupying70% of the collection space within the tube.

At this stage the filtrate collector 14 is sealed to the body of theblood separator device 8. Holding the device 8 vertically, open end up,a user introduces the inverted collection tube 10 and presses it gentlydown into the larger tubular body 12 of the separator device 8 to piercethe septum 10 a of the collection tube 10 with an opposed hypodermicneedle 20 that forms a capillary flow regulator or control element. Thedownward stroke of the collection tube 10 at first causes air only inthe closed volume below to be compressed. As shown on FIG. 8 thecollection tube 10 may travel to be fully inserted in the separatingdevice 8. But when the septum 10 a of the collection tube 10 reaches theprotruding hypodermic needle 20 and is pierced by it, pressure withinthe device 8 and the collection tube 10 is equilibrated.

For initiating filtering action, the Serum Collection Chamber (filtratecollector) 14 is partially then opened, permitting air to escape fromthe collector and bringing the region downstream from the filterassembly F to atmospheric pressure, thus creating a pressure differenceacross filter assembly F.

With this occurrence, air pressure above the blood within the collectiontube 10 becomes relatively higher than that below the Filter assembly F.This sets up a second automatic equilibrating action, in which thehigher air pressure in the collection tube 10 forces flow of blood outof the collection tube, downwardly through the hypodermic needle 20,into the space above the filter assembly F. In this implementation thepressure differential above atmospheric pressure thus drives bloodthrough the flow control and the filter media.

Preferably the compressed volume is small compared with the totaloriginal volume of the device. When the collection tube is pushed to itsstopped position, the “free” remaining volume of the device may be quitesmall.

The “free remaining volume” consists of the Serum Collection Chamber 14and the filter assembly F as well as the flow regulation assembly 20.

The established pressure differential is controlled by the Ideal GasLaw:

PV=Constant.

The initial conditions when the collection tube is about to beintroduced into the device P is atmospheric pressure.

Assuming that the inside diameter of the internal diameter of the mainbody is 11.0 mm at its open end is about equal to the diameter of thedeformable septum of the evacuated collection tube (Vacutainer™) suchthat the collection tube can be inserted without difficulty withalignment to its full length of 50.5 mm. The inner diameter of the mainbody 12 is slightly tapered such that it can easily be manufactured byinjection molding or otherwise. If the inside diameter of the main body,50 mm downward from the entry level is 10.5 mm, the volume of airdisplaced by the insertion of the collection tube is 4.58 cc.

The serum collection chamber is approximately 0.5 cc and the void volumeof the filter assembly approximately 1.0 cc with the pressure controland coupling region adding up to 0.75 cc, the total volume remainingadds to 2.25 cc.

The original air volume was 6.83 cc.

The “1.8 cc Vacutainer” has inner volume equal to 2.25 cc. and whenfilled with 1.8 cc of blood yields a void volume 0.45 cc.

The final volume of air is therefore 2.25+0.45=2.7 cc.

The Ideal Gas Law indicates that the pressure in the compressed deviceshall be:

1×6.83/2.7=2.5 atmosphere

This is the pressure of the air inside the blood collection tube.

When the “serum collection chamber” is opened to atmospheric pressurethe pressure differential propels blood out of the blood collectiontube.

Inside the Vacutainer the Ideal Gas Law applies. Prior to opening theserum collection chamber to atmospheric pressure, the conditions were:

P=2.5 atmosphere

V=0.45 cc

Opening the “serum collection chamber” to atmospheric pressure willbring that pressure to the inside of the Vacutainer and the air volumewill become:

V=1.125 cc

And approximately 0.675 cc of blood is forced out through the flowcontrol section, the filter and finally pushing the serum or plasma intothe serum collection chamber.

Approximately 0.25 cc of plasma is collected into the “serum collectionchamber”.

In respect of flow rate and flow geometry, the considerations andfindings described under the heading FLOW RATE AND FLOW GEOMETRY apply.

A number of embodiments of the invention have been described.Nevertheless, it will be understood that various modifications may bemade without departing from the spirit and scope of the invention. Forexample, the flow rate or pressure differential limiting control elementor device may take the form of a compliant tube wall section that tendsto expand outward to increase volume in response to pressure.Accordingly, other embodiments are within the scope of the followingclaims.

1. A filtering device for filtering blood to obtain serum or plasma in acontainer, the container having access at both ends, a filter locatedwithin the container, and a flow rate or pressure differential limitingcontrol element, the limiting element located upstream of the filter. 2.The filtering device of claim 1 in which the container is partiallyevacuated.
 3. The filtering device of claim 2 wherein the limitingcontrol element is located outside the container.
 4. The filteringdevice of claim 3 wherein the limiting control element is integral witha blood drawing needle assembly.
 5. (canceled)
 6. The filtering deviceof claim 1 wherein the container during operation is partiallypressurized.
 7. The filtering device of any of the foregoing claim 1wherein the limiting control element or device is located inside thecontainer.
 8. A filtering device, comprising a pre-evacuated container,a filter located within the container, and a flow rate or pressuredifferential limiting control element.
 9. The filtering device of claim8 wherein the control element comprises a pin hole or pin holes in aflow-blocking disk.
 10. The filtering device of claim 8 wherein thecontrol comprises a selected length of capillary tubing.
 11. Thefiltering device of claim 8 wherein the control element comprises a finemesh or porous foam.
 12. The filtering device of claim 8 wherein thecontrol element comprises a passage defined by a screw like segment. 13.A filtering device, comprising a pre-evacuated container, a filterlocated within the container, and a flow rate or pressure differentiallimiting control element is constructed to limit differential pressureacross the filter by restricting flow toward the filter, wherein thelimiting element is constructed to passively limit increase in thepressure differential rate of change across the filter to about 10 mmHg.14. The filtering device of claim 13 wherein it has a limiting controlelement or device preceding the filter that limits entering flow-rate ofwhole blood drawn from a living being.
 15. The filtering device of claim13 wherein it has a limiting control element or device preceding thefilter that defines entering pressure increase rate in whole blood drawnfrom a living being.
 16. The filtering device of claim 13 wherein it isportable or hand held, and includes a volume sized for blood drawn froma living being.
 17. The filtering device of claim 13 wherein thematerial of the filter comprises glass microfibers and micro-porousmembrane on a locating support.
 18. The filtering device of claim 13 inwhich the container is a tube.
 19. The filtering device of claim 13having an access septum at the inlet end of the container or tube. 20.The filtering device of claim 19 having an access septum at the outletend of the container or tube. 21-30. (canceled)
 31. The filtering deviceof claim 13 wherein the flow rate limiting control element is integralwith a blood drawing needle assembly. 32-33. (canceled)
 34. A method ofobtaining blood serum or plasma using the filtering device of claim 13.35-52. (canceled)
 53. A device comprising a pre-evacuated container, anda flow rate limiting control element limiting pressure and includingcapillary restriction for blood flow into the container and beingconstructed to provide said capillary restriction according to theHagen-Poiseuille law for a designed capillary radius, capillary lengthand pressure drop.
 54. The device of claim 53, wherein the pre-evacuatedcontainer includes a tube.
 55. The device of claim 54, wherein the tubeincludes inlet and outlet ends with access at both ends.